Xu W, Cooper G M
Division of Molecular Genetics, Dana-Farber Cancer Institute, Boston, Massachusetts 02115, USA.
Mol Cell Biol. 1995 Oct;15(10):5369-75. doi: 10.1128/MCB.15.10.5369.
The c-mos proto-oncogene is specifically expressed in female and male germ cells. Previous studies identified a negative regulatory element (NRE) upstream of the c-mos promoter that suppresses c-mos transcription in transfected NIH 3T3 cells. In this study, we used gel shift assays to detect proteins in nuclear extracts of NIH 3T3 cells that bind to the c-mos NRE in a sequence-specific manner. One protein was found to bind to a region of the NRE which was shown by site-directed mutagenesis to be required for suppression of c-mos transcription. This factor was present in nuclear extracts of several somatic cell lines and tissues but not in male germ cells in which c-mos is transcribed, suggesting that it is a somatic cell repressor of c-mos transcription. The binding site of the candidate repressor within the c-mos NRE consists of sequences related to putative NREs identified in two other male germ cell-specific genes (encoding protamine 2 and phosphoglycerate kinase 2). The c-mos repressor bound and could be UV cross-linked to these protamine 2 and phosphoglycerate kinase 2 gene sequences as a protein with an apparent molecular mass of approximately 30 kDa. The repressor binding site is also conserved in two other germ cell-specific genes (encoding testis-specific cytochrome c and heat shock-like protein 70), suggesting that the c-mos repressor may be generally involved in suppressing transcription of germ cell-specific genes in somatic cells.
原癌基因c-mos在雌性和雄性生殖细胞中特异性表达。先前的研究在c-mos启动子上游鉴定出一个负调控元件(NRE),该元件在转染的NIH 3T3细胞中抑制c-mos转录。在本研究中,我们使用凝胶迁移试验来检测NIH 3T3细胞核提取物中以序列特异性方式与c-mos NRE结合的蛋白质。发现一种蛋白质与NRE的一个区域结合,定点诱变表明该区域是抑制c-mos转录所必需的。这种因子存在于几种体细胞系和组织的核提取物中,但在转录c-mos的雄性生殖细胞中不存在,这表明它是c-mos转录的体细胞抑制因子。c-mos NRE内候选抑制因子的结合位点由与另外两个雄性生殖细胞特异性基因(编码鱼精蛋白2和磷酸甘油酸激酶2)中鉴定的推定NRE相关的序列组成。c-mos抑制因子与这些鱼精蛋白2和磷酸甘油酸激酶2基因序列结合,并可通过紫外线交联形成一种表观分子量约为30 kDa的蛋白质。抑制因子结合位点在另外两个生殖细胞特异性基因(编码睾丸特异性细胞色素c和热休克样蛋白70)中也保守,这表明c-mos抑制因子可能普遍参与抑制体细胞中生殖细胞特异性基因的转录。